Abstract
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is an emerging technique for the characterization of biological systems. With the development of novel ion sources such as cluster ion beams, ionization efficiency has been increased, allowing for greater amounts of information to be obtained from the sample of interest. This enables the plotting of the distribution of chemical compounds against position with submicrometer resolution, yielding a chemical map of the material. In addition, by combining imaging with molecular depth profiling, a complete 3-dimensional rendering of the object is possible. The study of single biological cells presents significant challenges due to the fundamental complexity associated with any biological material. Sample preparation is of critical importance in controlling this complexity, owing to the fragile nature of biological cells and to the need to characterize them in their native state, free of chemical or physical changes. Here, we describe the four most widely used sample preparation methods for cellular imaging using ToF-SIMS, and provide guidance for data collection and analysis procedures.
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References
Roddy TP, Cannon DM, Meserole CA, Winograd N, Ewing AG (2002) Imaging of freeze-fractured cells with in situ fluorescence and time-of-flight secondary ion mass spectrometry. Anal Chem 74(16):4011–4019
Piehowski PD, Kurczy ME, Willingham D, Parry S, Heien ML, Winograd N, Ewing AG (2008) Freeze-etching and vapor matrix deposition for ToF-SIMS imaging of single cells. Langmuir 24(15):7906–7911
Vickerman JC (2011) Molecular imaging and depth profiling by mass spectrometry-SIMS, MALDI or DESI? Analyst 136(11):2199–2217
Berman ESF, Fortson SL, Checchi KD, Wu L, Felton JS, Wu KJJ, Kulp KS (2008) Preparation of single cells for imaging/profiling mass spectrometry. J Am Soc Mass Spectrom 19(8):1230–1236
Fletcher JS, Rabbani S, Henderson A, Lockyer NP, Vickerman JC (2011) Three-dimensional mass spectral imaging of HeLa-M cells: sample preparation, data interpretation and visualisation. Rapid Commun Mass Spectrom 25(7):925–932
Malm J, Giannaras D, Riehle MO, Gadegaard N, Sjovall P (2009) Fixation and drying protocols for the preparation of cell samples for time-of-flight secondary Ion mass spectrometry analysis. Anal Chem 81(17):7197–7205
Rabbani S, Fletcher JS, Lockyer NP, Vickerman JC (2011) Exploring subcellular imaging on the buncher-ToF J105 3D chemical imager. Surf Interface Anal 43(1–2):380–384
Parry S, Winograd N (2005) High-resolution TOF-SIMS imaging of eukaryotic cells preserved in a trehalose matrix. Anal Chem 77(24):7950–7957
Hoppert M, Holzenburg A (1998) Electron microscopy in microbiology. BIOS Scientific Publishers, Oxford
Severs NJ, Newman TM, Shotton DM (1995) A practical introduction to rapid freezing techniques. In: Severs NJ, Shotton DM (eds) Rapid freezing, freeze fracture and deep etching. Wiley-Liss, New York, NY
Lanekoff I, Kurczy ME, Adams KL, Malm J, Karlsson R, Sjovall P, Ewing AG (2011) An in situ fracture device to image lipids in single cells using ToF-SIMS. Surf Interface Anal 43(1–2):257–260
Brison J, Benoit DSW, Muramoto S, Robinson M, Stayton PS, Castner DG (2011) ToF-SIMS imaging and depth profiling of HeLa cells treated with bromodeoxyuridine. Surf Interface Anal 43(1–2):354–357
Piwowar AM, Keskin S, Delgado MO, Shen K, Hue JJ, Lanekoff I, Ewing AG, Winograd N (2013) C60-ToF SIMS imaging of frozen hydrated HeLa cells. Surf Interface Anal 45(1):302–304
Piwowar AM, Fletcher JS, Kordys J, Lockyer NP, Winograd N, Vickerman JC (2010) Effects of cryogenic sample analysis on molecular depth profiles with TOF-secondary ion mass spectrometry. Anal Chem 82(19):8291–8299
Roddy TP, Cannon DM, Ostrowski SG, Ewing AG, Winograd N (2003) Proton transfer in time-of-flight secondary ion mass spectrometry studies of frozen-hydrated dipalmitoylphosphatidylcholine. Anal Chem 75(16):4087–4094
Braun RM, Blenkinsopp P, Mullock SJ, Corlett C, Willey KF, Vickerman JC, Winograd N (1998) Performance characteristics of a chemical imaging time-of-flight mass spectrometer. Rapid Commun Mass Spectrom 12(18):1246–1252
Fletcher JS, Lockyer NP, Vaidyanathan S, Vickerman JC (2007) TOF-SIMS 3D biomolecular imaging of Xenopus laevis oocytes using buckminsterfullerene (C-60) primary ions. Anal Chem 79(6):2199–2206
Stryer L (1981) Biochemistry, 2nd edn. W. H. Freeman and Company, San Francisco, CA
Acknowledgments
This project was supported by grants from the National Center for Research Resources (5P41RR031461) and the National Institute of General Medical Sciences (8 P41 GM103391) from the National Institutes of Health. In addition, infrastructure support from the National Science Foundation under grant number CHE-0908226 and by the Division of Chemical Sciences at the Department of Energy grant number DE-FG02-06ER15803 is acknowledged.
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Winograd, N., Bloom, A. (2015). Sample Preparation for 3D SIMS Chemical Imaging of Cells. In: He, L. (eds) Mass Spectrometry Imaging of Small Molecules. Methods in Molecular Biology, vol 1203. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1357-2_2
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DOI: https://doi.org/10.1007/978-1-4939-1357-2_2
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